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71377 NoShift™ Transcription Factor Assay Kit

71377
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          Description
          OverviewThe NoShift™ Transcription Factor Assay Kit is a microassay plate–based method for analysis of DNA-binding proteins that alleviates many of the problems associated with gel shift assays. The gel shift, or electrophoretic mobility shift assay (EMSA), is a method for analyzing interactions between proteins and DNA. This technique, first published in 1981 (Fried 1981, Garner 1981), is based on the reduced electrophoretic mobility of protein–DNA complexes compared with that of DNA alone under nondenaturing conditions. The traditional gel shift assay is tedious and time consuming, requires radio-labeled probe, and often requires extensive optimization. The 96-well NoShift plate format is versatile and enables simultaneous analysis of multiple binding factors in less than five hours without the use of radioisotopes. A biotinylated oligonucleotide is mixed with a protein sample and the protein–DNA complex is captured on streptavidin plates. The bound transcription factor is detected with a specific antibody followed by a second antibody–HRP conjugate and chromogenic substrate.

          Each NoShift Transcription Factor Assay Kit comes with one 96-strip-well streptavidin-coated plate, binding reaction buffer, antibody dilution buffer, plate wash buffer, and TMB substrate and provides enough reagents for 100 reactions. The NoShift Kit is designed for use with oligonucleotides defining a protein binding consensus sequence provided by the purchaser and an antibody to a DNA binding protein.

          Catalogue Number71377
          Brand Family Novagen®
          References
          ReferencesFried, M. and Crothers, D.M. 1981. Nucleic Acids Res. 9, 6505. Garner, M.M. and Revzin, A. 1981. Nucleic Acids Res. 9, 3047.
          Product Information
          4 × 1 ml4X NoShift™ Bind Buffer
          150 µlSalmon Sperm DNA
          125 µlPoly(dI:dC):Poly(dI:dC)
          30 ml10X NoShift Wash Buffer
          20 mlNoShift Antibody Dilution Buffer
          1 eaStreptavidin Plate
          12 mlTMB Substrate
          100 µl100 mM DTT
          3 eaAluminum Plate Sealer
          Quality LevelMQ100
          Applications
          Biological Information
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          Materials Information
          Toxicological Information
          Safety Information according to GHS
          Safety Information
          Product Usage Statements
          Storage and Shipping Information
          Ship Code Blue Ice Only
          Toxicity Multiple Toxicity Values, refer to MSDS
          Storage Multiple storage requirements
          Do not freeze Ok to freeze
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          Supplemental Information
          Specifications

          Documentation

          NoShift™ Transcription Factor Assay Kit Certificates of Analysis

          TitleLot Number
          71377

          References

          Reference overview
          Fried, M. and Crothers, D.M. 1981. Nucleic Acids Res. 9, 6505. Garner, M.M. and Revzin, A. 1981. Nucleic Acids Res. 9, 3047.

          Citations

          Title
        • Markus Bredel, et al. (2006) Tumor necrosis factor-α-induced protein 3 as a putative regulator of nuclear factor-κB-mediated resistance to O6-alkylating agents in human glioblastomas. Journal of Clinical Oncology 24, 274-287.
        • Zachary T. Resch, et al. (2005) Stress-activated signaling pathways mediate the stimulation of pregnancy-associated plasma protein-A expression in cultured human fibroblasts. Endocrinology 147, 885-890.
        • Andrei P. Pomerantsev, Olga M. Pomerantsev and Stephen H. Leppla. (2004) A spontaneous translational fusion of Bacillus cereus plcR and papR activates transcription of plcR-dependent genes in Bacillus anthracis via binding with a specific pallindromic sequence. Infection and Immunity 72, 5814-5823.
        • User Protocols

          Title
          TB403 NoShift™ Transcription Factor Assay Kit